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Ablation of Papillar Cell Function in Brassica Flowers Results in the Loss of Stigma Receptivity to Pollination.

Plant reproduction in crucifers is dependent on interactions that occur at the stigma surface between the male gametophyte (pollen and pollen tube) and papillar cells. To dissect these complex interactions, papillar cells were genetically ablated by targeting the expression of a toxin to appropriate cells of the flower with a flower-specific and developmentally regulated promoter. In transgenic Brassica plants that expressed the toxic gene fusion, flower morphology was normal except for aberrant papillar cell development and partial pollen sterility. Microscopic, biochemical, and functional analyses, mainly focused on papillar cell responses, revealed that papillar cells lost their ability to elongate, to synthesize cell-specific proteins, and to support pollen germination after self- or cross-pollination. This loss of stigma receptivity to pollination was mimicked by treating pistils with protein phosphatase inhibitors. Differences in the effects of genetic and chemical ablation on the pollination responses of Brassica and Arabidopsis flowers are discussed and are ascribed in part to a requirement for phosphorylation/dephosphorylation events in Brassica but not in Arabidopsis.

Journal Article↗

Pollination- or Wound-Induced Kaempferol Accumulation in Petunia Stigmas Enhances Seed Production.

Flavonols are essential for pollen germination and tube growth in petunia and can be supplied by either the pollen or stigma at pollination. HPLC analysis and a sensitive bioassay demonstrated that both pollination and wounding induce flavonol accumulation, especially kaempferol, in the outer cell layers and exudate of the stigma. Pollination and wounding induced nearly identical flavonol kinetics and patterns of accumulation in the same target tissue, suggesting that they share elements of a common signal transduction pathway. The wound response was systemic, because kaempferol accumulated in the stigma when distal tissues, such as the corolla, stamens, or sepals, were wounded. We have exploited the germination requirement for flavonols and the high level of kaempferol that accumulates after wounding to enhance plant fecundity. Seed set was significantly increased by mechanically wounding the corolla and stamens prior to the application of pollen to the stigma. A reproductive role for a plant secondary metabolite and the specific function of stigmatic kaempferol are discussed from an evolutionary perspective.

Journal Article↗

Phenotypic selection in hawkmoth-pollinated Platanthera bifolia: targets and fitness surfaces.

The present study explored phenotypic selection on phenological and morphological reproductive traits in hawkmoth-pollinated Platanthera bifolia (Orchidaceae), a Eurasian perennial herb displaying bisexual, long-spurred flowers. The work was carried out during three flowering seasons (1993-1995) in a Swedish population. Fitness was estimated as the number of pollinia removed (male fitness) and fruits produced (female fitness). Targets and patterns of selection were compared between years and sex functions by the use of multiple linear regression (including correlational selection estimates, i.e., of combination of traits), analysis of covariance, and projection pursuit regression (PPR). Results from the nonparametric surface-fitting-method PPR showed that selection was mostly linear, thus justifying the use of the parametric methods. In all study years, male and female fitness were highest in plants with many flowers. This reflects that flower number sets an upper limit to fitness and that a large influorescence attracts more pollinators. In 1994, the summer was dry and the average spur length of P. bifolia was shorter than in the other years. In this year, male and female fitness were positively related to spur length, apparently because the spur of short-spurred plants was somewhat too short relative to the tongue length of the local pollinator for optimal pollen export and import. Additionally, the dry weather in 1994 caused a tendency for correlational selection, which was not found in the other years of study. Among small individuals (apparently more sensitive to drought than large ones), early-flowering plants had higher male and female fitness. The results show that patterns of selection may vary both between years and between sex functions in perennial hermaphroditic plants. The present study is one of the first to consider correlational selection in plants, which probably is of common occurrence and deserves to be investigated more.

Animals↗

Consequences of floral complexity for bumblebee-mediated geitonogamous self-pollination in Salvia nipponica Miq. (Labiatae).

I address how floral complexity influences geitonogamous self-pollination through manipulation of pollinator behavior in Salvia nipponica. The pivoting stamens of S. nipponica hinder nectar-collecting bumblebees from crawling into flowers, increasing the probing time per flower. I predicted that longer probing times would reduce the relative cost of moving between plants, causing bees to leave plants earlier. To test this prediction, I simplified S. nipponica flowers by removing the stamens from all open flowers within a 75-m2 quadrat. Bumblebees probed these flowers more quickly than intact flowers, but the stamen removal affected neither the frequency of flower revisitation nor the flight distance between plants. In response to the decrease in the probing time per flower, bees probed more flowers on these plants. Therefore, in S. nipponica, floral complexity reduces the opportunity for geitonogamous self-pollination. Stamen removal also increased bee visitation per flower, suggesting that this sort of complexity deters visitation. To keep complex flowers attractive, therefore, selection might increase floral rewards or longevity. Floral complexity might evolve in an integrative manner with the rest of the floral phenotype.

Animals↗

Phylogeography of pollinator-specific sexually deceptive Chiloglottis taxa (Orchidaceae): evidence for sympatric divergence?

Divergence in sexually deceptive orchids is thought to occur through shifts in the attraction of specific pollinators, a process that is mediated by changes in the floral odours that lure sexually excited male insects. We investigated the origin of reproductive isolation in a sexually deceptive species complex of Chiloglottis R.Br. (Orchidaceae: Diurideae). Two geographically separated montane regions in eastern Australia were sampled, each containing sympatric pairs of orchid taxa presently found under the name, Chiloglottis pluricallata. Behavioural tests confirmed at least three distinct orchid taxa that specifically attract different pollinators. An artificial crossing experiment among two taxa from one region demonstrated their interfertility, and confirmed isolation to be a function of pollinator attraction. A phylogeographic analysis using amplified fragment length polymorphisms (AFLPs) indicated that samples from each geographical region are most closely related, a pattern consistent with in situ or sympatric divergence. However, an extensive population genetic study on two taxa from one region failed to entirely reject the possibility of intertaxon gene flow. Although clear genetic differentiation of the taxa is evident in two out of three sites where both grow in direct sympatry, overall, the two taxa are not strongly distinguished by AFLP markers. The reconstruction of a simple bifurcating pattern of divergence may be confounded by a combination of contemporary population-level processes operating within each taxon, the retention of ancestral polymorphism or intertaxon gene flow.

Alleles↗

Environment and pollinator-mediated selection on parapatric floral races of Mimulus aurantiacus.

We tested whether selection by pollinators could explain the parapatric distribution of coastal red- and inland yellow-flowered races of Mimulus aurantiacus (Phrymaceae) by examining visitation to natural and experimental populations. As a first step in evaluating whether indirect selection might explain floral divergence, we also tested for local adaptation in early life stages using a reciprocal transplant experiment. Hummingbirds visited flowers of each race at similar rates in natural populations but showed strong (>95%) preference for red flowers in all habitats in experimental arrays. Hawkmoths demonstrated nearly exclusive (>99% of visits) preference for yellow flowers and only visited in inland regions. Strong preferences for alternative floral forms support a direct role for pollinators in floral divergence. Despite these preferences, measures of plant performance across environments showed that red-flowered plants consistently survived better, grew larger and received more overall pollinator visits than yellow-flowered plants. Unmeasured components of fitness may favour the yellow race in inland habitats. Alternatively, we document a marked recent increase in inland hummingbird density that may have caused a change in the selective environment, favouring the eastward advance of red-flowered plants.

Animals↗

Pollination of Ravenala madagascariensis and Parkia madagascariensis by Eulemur macaco in Madagascar.

Primates are known to be important in dispersal of seeds of tropical rainforest trees, but their role in pollination is very poorly documented. Although the 'traveller's palm' Ravenala madagascariensis is widespread, only a single well-documented report exists for its pollination by Malagasy prosimians. Black lemurs, Eulemur macaco, exploit nectaries of Parkia as well as Ravenala systematically at the massifs of Ambato and Lokobe, and almost certainly contribute substantially to their pollination, confirming a proposal first made by Sussman and Raven [Science 1978;200: 731-736].

Journal Article↗

Deep mtDNA divergences indicate cryptic species in a fig-pollinating wasp.

BACKGROUND: Figs and fig-pollinating wasps are obligate mutualists that have coevolved for ca. 90 million years. They have radiated together, but do not show strict cospeciation. In particular, it is now clear that many fig species host two wasp species, so there is more wasp speciation than fig speciation. However, little is known about how fig wasps speciate. RESULTS: We studied variation in 71 fig-pollinating wasps from across the large geographic range of Ficus rubiginosa in Australia. All wasps sampled belong to one morphological species (Pleistodontes imperialis), but we found four deep mtDNA clades that differed from each other by 9-17% nucleotides. As these genetic distances exceed those normally found within species and overlap those (10-26%) found between morphologically distinct Pleistodontes species, they strongly suggest cryptic fig wasp species. mtDNA clade diversity declines from all four present in Northern Queensland to just one in Sydney, near the southern range limit. However, at most sites multiple clades coexist and can be found in the same tree or even the same fig fruit and there is no evidence for parallel sub-division of the host fig species. Both mtDNA data and sequences from two nuclear genes support the monophyly of the "P. imperialis complex" relative to other Pleistodontes species, suggesting that fig wasp divergence has occurred without any host plant shift. Wasps in clade 3 were infected by a single strain (W1) of Wolbachia bacteria, while those in other clades carried a double infection (W2+W3) of two other strains. CONCLUSION: Our study indicates that cryptic fig-pollinating wasp species have developed on a single host plant species, without the involvement of host plant shifts, or parallel host plant divergence. Despite extensive evidence for coevolution between figs and fig wasps, wasp speciation may not always be linked strongly with fig speciation.

Animals↗

Pollen-stigma interactions in Brassica oleracea. II. The fate of stigma surface proteins following pollination and their rôle in the self-incompatibility response.

Mature, self-incompatible stigmas exposed to cycloheximide for 2 h prior to pollination supported identical germination and growth of both cross and self pollen. Treatment of self-pollinated pistils with cycloheximide resulted in the germination of hitherto inactive pollen after some 2-4 h. Pollen germination and initial tube growth in an in vitro germination medium were not significantly affected by cycloheximide. A continuous synthesis of stigmatic proteins is therefore essential for the operation of the self-incompatibility (S.I.) system. However, light-microscope autoradiography of stigmas fed with L-[3H]leucine prior to pollination revealed no movement of stigmatic proteins into the pollen, independent of the compatibility of the pollen with respect to the stigma. Further, tunicamycin, when applied in the same way as cycloheximide, had no effect on the S.I. system. These results are discussed in terms of the proposed cycling of proteins in the papillar cell wall and the involvement of a stigmatic glycoprotein in the S.I. response.

Brassica↗

Echoes of bat-pollinated bell-shaped flowers: conspicuous for nectar-feeding bats?

Nectar-feeding glossophagine bats searching for flowers are guided by their echolocation system as well as olfactory cues in detecting and recognizing nectar sources. Therefore, chiropterophilous plants, which depend on these bats as pollinators, may be expected to have evolved acoustically conspicuous flowers that facilitate detection. As it is poorly understood how bats manage to find and recognize flowers acoustically, we investigated the echoes of some of the flowers pollinated by bats. Echoes of bell-shaped bat-pollinated flowers have characteristic features with respect to the echoes they reflect to a calling bat and differ from the echoes of leaves or other objects in their surroundings: the echoes are comparatively long and of complex spectral composition. Owing to the specific shape of the flowers, characteristic 'spectral directional patterns' result when the spectra of the echoes are plotted against the angle of sound incidence. We suggest that bats are able to recognize such flowers - and probably other objects as well - not only by a characteristic spectral composition of the echo but also by comparing sequential echoes, at the same time taking into account their exact calling position relative to the object.

Adaptation, Biological↗

Ants visit nectaries of Epidendrum denticulatum (Orchidaceae) in a Brazilian rainforest: effects on herbivory and pollination.

Epidendrum denticulatum (Orchidaceae) produces nectar on the petioles of buds, flowers, and fruits (extrafloral nectaries) but no nectar is found on its flowers, and it is probably a deceptive species. In the Brazilian Atlantic rainforest, some aspects of both the ecology and behavior of Camponotus sericeiventris (Formicinae) and Ectatomma tuberculatum (Ponerinae), two ant species foraging on E. denticulatum extrafloral nectaries, were investigated. Both experiments, using termites as baits and field observations, suggest that these ant species are able to prevent reproductive organ herbivory, without affecting pollinator behaviour. Since a low fruit set is often cited as a characteristic of the family, especially for deceptive species, ants attracted to orchid inflorescences protect reproductive structures and increase the probability of pollination success. Epidendrum denticulatum flowers were visited and probably pollinated by Heliconius erato (Nymphalidae) and Euphyes leptosema (Hesperiidae).

Animals↗

Alfalfa leafcutting bee population dynamics, flower availability, and pollination rates in two Oregon alfalfa fields.

Since the 1970s, it has become increasingly difficult for U.S. alfalfa seed producers to maintain Megachile rotundata (F.) populations used for alfalfa, Medicago sativa L., pollination. In 1998, we monitored M. rotundata population dynamics and foraging behavior, as well as alfalfa bloom and pollination rates in two fields in eastern Oregon. Despite marked differences in bee management, establishment was very similar in the two fields (approximately 0.5 females per nesting cavity) and lagged peak bloom by approximately 2 wk. Pollination rates increased from 0-10% in the first 3 wk to 80-90% in week 4-5. By then, M. rotundata females had difficulty finding untripped (nonpollinated) flowers and visited large numbers of already tripped or not fully matured flowers. M. rotundata progeny mortality was very high (54-78%). Estimated seed yields were similar in both fields. We contend similar seed yields, and improved bee production, could be accomplished with smaller bee populations, better timed with alfalfa bloom.

Animals↗

Effects of honey bee (Hymenoptera: Apidae) and bumble bee (Hymenoptera: Apidae) presence on cranberry (Ericales: Ericaceae) pollination.

Honey bees, Apis mellifera L., are frequently used to pollinate commercial cranberries, Vaccinium macrocarpon Ait., but information is lacking on the relative contribution of honey bees and native bees, the effects of surrounding vegetation on bee visitation, and on optimal timing for honey bee introduction. We begin with a descriptive study of numbers of honey bees, bumble bees, and other bees visiting cranberry blossoms, and their subsequent effect on cranberry yield, on three cranberry properties in 1999. The property surrounded by agricultural land, as opposed to wetlands and woodlands, had fewer numbers of all bee types. In 2000, one property did not introduce honey bee colonies, providing an opportunity to document the effect of lack of honey bees on yield. With no honey bees, plants along the edge of the bed had significantly higher berry weights compared with nonedge plants, suggesting that wild pollinators were only effective along the edge. Comparing the same bed between 1999, with three honey bee colonies per acre, and 2000, with no honey bees, we found a significant reduction in average berry size. In 2000, we compared stigma loading on properties with and without honey bees. Significantly more stigmas received the minimum number of tetrads required for fruit set on the property with honey bees. Significantly more tetrads were deposited during mid-bloom compared with early bloom, indicating that mid-bloom was the best time to have honey bees present. This study emphasizes the importance and effectiveness of honey bees as pollinators of commercial size cranberry plantings.

Animals↗

Pollination of Ipomopsis aggregata (Polemoniaceae): effects of intra- vs. interspecific competition.

Although plants may simultaneously experience intra- and interspecific competition for pollination, their relative strength has rarely been experimentally evaluated. Yet because intra- and interspecific competition can be caused by different mechanisms, their effect on the ecology and evolution of plants may differ. To determine the relative strength of intra- and interspecific competition for pollination, I manipulated the presence of heterospecifics and density of conspecifics using Ipomopsis aggregata as the focal species. All plots contained I. aggregata and Castilleja linariaefolia, but C. linariaefolia inflorescences were removed from half of the plots to create the heterospecifics-absent treatment. Within each plot, all I. aggregata inflorescences were removed from a 5-m radius around a focal plant to create a low conspecific density experimental unit, and a group of 12 I. aggregata plants/1 m(2) was designated as a high conspecific density unit. Conspecific pollen deposition was reduced when C. linariaefolia was present but was not influenced by I. aggregata density. Although seed set per fruit was reduced by 17% when C. linariaefolia was present, it was not significantly influenced by either treatment. Interspecific competition for pollination is stronger than intraspecific competition in the I. aggregata-C. linariaefolia system, but neither process appears to influence plant fitness.

Journal Article↗

[Floral biology and pollinators of Trichocereus pasacana (Cactaceae) in Parque Nacional Los Cardones, Argentina].

Many columnar cacti are bat pollinated. It has been suggested that this kind of pollination would be more important in tropical than in temperate regions where flowers are open only one night. Thrichocereus pasacana produces big and resistant white flowers. We analyzed flower characteristics, floral cycle, stigmatic receptivity, nectar production, pollen presence and floral visitors in a T. pasacana population at National Park Los Cardones (Salta, Argentina) in November 1997. Flower features were constant between individuals of the population. Flowers start opening at evening and anthesis time is from 18 to 40 hs. The estigma was receptive throughout the floral cycle. Anther dehiscence occurs with flower opening. Nectar production was highest between 18 to 24 hs. Although T. pasacana are open during the night, floral visitors are diurnal. The most frequent was Xylocopa sp. In the study area, nectarivorous bats were not detected. The morphological features of T. pasacana flowers were similar but bigger compared to other columnar cacti. Anthesis time was also longer while nectar production was lower. T. pasacana pollination at National Park Los Cardones is done by bees.

Animals↗

Oriental orchid (Cymbidium pumilum) attracts drones of the Japanese honeybee (Apis cerana japonica) as pollinators.

The discovery that drones of the Japanese honeybee (Apis cerana japonica) pollinate the oriental orchid (Cymbidium pumilum) is reported. Drones are attracted to the orchid flower aroma mainly during their mating flights in April through May. Some drones cluster on the flower racemes and others insert their heads deep into the flowers. Drones with pollinia on their scutellum visit other orchids, which facilitates pollination. Individual workers and swarming colonies are also strongly attracted by the flower aroma, but the allopatric western honeybee (Apis mellifera) is not attracted.

Animals↗

Molecular studies on genetic integrity of open-pollinating species rye (Secale cereale L.) after long-term genebank maintenance.

The genetic integrity of six accessions represented by 14 sub-populations of the open-pollinating species rye ( Secale cereale L.) was investigated. Seeds available from a herbarium collection (first regeneration) and from the cold store (most recent regeneration) were multiplied two to fourteen times and fingerprinted using microsatellite markers. Four accessions had significantly different allele frequencies. These were multiplied seven to thirteen times. Nearly 50% of the alleles discovered in the original samples were not found in the material present in the cold store. However alleles were detected in the most recently propagated sub-populations, that were not observed in the investigated plants of the original one. The change in allele frequencies is a continuous process. Reasons for the occurrence of genetic changes and consequences for managing open pollinating species maintained in ex situ genebanks are discussed.

Gene Frequency↗

Field metabolic rates of phytophagous bats: do pollination strategies of plants make life of nectar-feeders spin faster?

Recently, it was argued that extrinsic factors, such as high foraging costs, lead to elevated field metabolic rates (FMR). We tested this suggestion by comparing the FMR of nectar-feeding and fruit-eating bats. We hypothesized that the foraging effort per energy reward is higher for nectar-feeding mammals than for fruit-eating mammals, since energy rewards at flowering plants are smaller than those at fruiting plants. Using the doubly labelled water method, we measured the FMR of nectar-feeding Glossophaga commissarisi and fruit-eating Carollia brevicauda, which coexisted in the same rainforest habitat and shared the same daytime roosts. Mass-specific FMR of G. commissarisi exceeded that of C. brevicauda by a factor of almost two: 5.3+/-0.6 kJ g(-1) day(-1) for G. commissarisi and 2.8+/-0.4 kJ g(-1) day(-1) for C. brevicauda. Since nectar-feeding bats imbibe nectar droplets of only 193 J energy content during each flower visit, a G. commissarisi bat has to perform several 100 flower visits per night to meet its energy requirement. The fruit-eating C. brevicauda, on the other hand, needs to harvest only 3-12 Piper infructescenses per night, as the energy reward per Piper equals ca. 6-30 kJ. We argue that the flowering and fruiting plants exert different selective forces on the foraging behaviour and energetics of pollinators and the seed dispersers, respectively. A comparison between nectar-feeding and non-nectar-feeding species in various vertebrate taxa demonstrates that pollinators have elevated FMRs.

Animals↗